Regulator of G protein signaling

RGS proteins

Regulator of G-proteins signaling (RGS) proteins play a critical role in many G protein-dependent signaling pathways. Thus, RGS proteins have been implicated in a wide range of pathologies, including cancer, hypertension, arrhythmias, drug abuse and schizophrenia. RGS proteins ‘turn off’ heterotrimeric (αβγ) G-proteins and thereby determine the duration of G protein–mediated signaling events. Therefore, RGS proteins function as GTPase Activating Proteins (GAPs), and this GAP activity is mediated by allosteric interactions. RGS proteins are selective for binding to the transition state of Gα(GTP → GDP + P_i), which can be mimicked by Gα-GDP bound with the planar ion aluminum tetrafluoride(AlF_-4).

Like many signaling proteins, RGS proteins comprise a large and diverse family. In human genome, Thirty-seven RGS proteins are encoded by gene loci; this collection of related proteins has been divided into 10 different subfamilies based on the relatedness of their RGS domain sequence and their multiple domain architectures. About 20 ‘canonical’ RGS proteins can in theory downregulate any of the 16 activated Gα subunits, although in practice they interact only with members of the G_i and G_q families. In these proteins, the ~120-residue RGS homology domain functions as a GTPase-activating protein (GAP) for GTP-bound Gα subunits.

Phylogenetic tree of 19 human RGS domains. RGS proteins whose activity was tested are colored by their GAP activity, RGS proteins with high GAP activity (green), RGS proteins with low but discernible activities (purple) and RGS2 had no measurable activity (red). ^[1]

In addition to these domains, diverse proteins subfamilies that include the ~120-residue RGS homology domain bear additional protein-protein interaction domains beyond their signature RGS domain with Gα GAP activity. R7-subfamily members share a multi-domain protein architecture composed of DEP and GGL domains on the N-terminal side of the RGS domain. R12-subfamily members possess a tandem repeat of Ras binding domains (RBDs) and a single GoLoco motif.

Heterotrimeric G-proteins family

Human heterotrimeric G-proteins are derived from 35 genes: 16 encoding α subunits, 5 β and 14 γ subunits. The α subunits function as guanine nucleotide on-off switches, mechanistically similar to other G-proteins that are enzymatic GTPases. G-proteins interact with diverse protein partners, such as G-protein coupled receptors (GPCRs), downstream effectors, and other proteins.

One important G-protein interaction is with members of the RGS protein family. This interaction occurs when the G-protein alpha subunit is activated, and depends on the Gα class, which in turn depends on their sequence that classifies them into several sub-types.

Phylogenetic tree of mammalian G-protein α-subunits classified to 4 groups based on there sequence identity.^[2] Gα_i and Gα_q families have selectivity towards The RGS members of R4-subfamily and R12-subfamily, Gα_z subunits were suggested to have selectivity for the RGS members of RZ-subfamily . The other two subfamilies Gα_s and Gα_12,13 might doing interaction with diverse proteins subfamilies that include the ~120-residue RGS homology domain but can't interact with canonical RGS members.

Structural highlights

1AGR is a complex of RGS4 and Gα_i1 proteins defined by X-ray crystallographic analysis. corresponds to an array of nine α-helices that fold into two small subdomains, both subdomains are required for it's GAP activity. α1, α2, α3, α4, α5 and α6 helices are colored in blue, aqua, yellow, coral, magenta and dark green respectively while helices α7, α8 and α9 are colored red.

Gα_i1 subunits adopt a conserved fold composed of , a helical domain of six α helices shown as blue cartoon and a GTPase domain shown in gray cartoons. The GTPase domain hydrolyzes GTP and provides most of Gα's binding surfaces for Gβγ, receptors, effectors and RGS proteins. contains three flexible regions designated switch-I presented as blue sticks, switch-II presented as magenta sticks and switch-III presented as green sticks that change conformation in response to GTP binding and hydrolysis, GDP–Mg⁺², bound in the active site of Gα_i1 is shown as a ball-and-stick model. The three switch regions of Gα_i1: residues 176–184, 201–215, and 233–241, respectively . ^[3]

RGS-G proteins interactions

There are many RGS protein residues in the vicinity of the RGS domain–Gα interface that contribute to RGS-G proteins interaction. These residues classified into two major groups. First group is Significant & Conserved residues that located mainly in the center of the RGS domain–Gα interface and have the primary role in accelerating Gα GTPase by stabilizing Gα in a conformation optimal for GTP hydrolysis. Whereas the second group is putative Modulatory residues that located mostly at the periphery of this interface where they contribute to Gα subunit recognition.^[4]

Gα subunits participate in a range of interactions with a variety of other proteins. Therefore, they have interfaces than interact selectively with receptors, effector subfamilies and RGS proteins. However the Gα residues that interact specifically with RGS proteins are highly conserved. Gα Residues located on the three Gα switch regions interact with Significant & Conserved RGS residues. This makes sense because of the pivotal role of the switch regions in GTP hydrolysis that is catalyzed by RGS proteins. On the other hand, Gα residues located in switch regions II and III and multiple residues in the Gα all-helical domain interact with Modulatory RGS residues.